Split Green Fluorescent Protein–Based Contact Site Sensor (SPLICS) for Heterotypic Organelle Juxtaposition as Applied to ER–Mitochondria Proximities
| Autor: | Marisa Brini, Tito Calì |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Cell physiology Endosome Chemistry Split GFP Endoplasmic reticulum ER–Mitochondria tethering Mitochondria Organelle contact sites SPLICS Cellular homeostasis Golgi apparatus Peroxisome Cell biology 03 medical and health sciences symbols.namesake 030104 developmental biology 0302 clinical medicine Lipid droplet Organelle symbols 030217 neurology & neurosurgery |
| Zdroj: | Methods in Molecular Biology ISBN: 9781071612613 |
| DOI: | 10.1007/978-1-0716-1262-0_23 |
| Popis: | In the last decades, membrane contact sites (MCSs) have been the object of intense investigation in different fields of cell physiology and pathology and their importance for the correct functioning of the cell is now widely recognized. MCS between any known intercellular organelles, including endoplasmic reticulum (ER), mitochondria, Golgi, endosomes, peroxisomes, lysosomes, lipid droplets, and the plasma membrane (PM), have been largely documented and in some cases the molecules responsible for the tethering also identified. They represent specific membrane hubs where a tightly coordinated exchange of ions, lipids, nutrients, and factors required to maintain proper cellular homeostasis takes place. Their delicate, dynamic, and sometimes elusive nature prevented and/or delayed the development of tools to easily image interorganelle proximity under physiological conditions and in living organisms. Nowadays, this aspect received great momentum due to the finding that MCSs' dysregulation is involved in several pathological conditions. We have recently developed modular, split-GFP-based contact site sensors (SPLICS) engineered to fluoresce when homo- and heterotypic juxtapositions between ER and mitochondria occur over a range of specific distances. Here we describe in detail, by highlighting strengths and weaknesses, the use and the application of these novel genetically encoded SPLICS sensors and how to properly quantify short- and long-range ER-mitochondria interactions. |
| Databáze: | OpenAIRE |
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